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目的：评价葡萄籽原花青素对过氧化氢导致的人晶状体上皮细胞损伤的保护作用。 方法：对人晶状体上皮细胞进行培养，分别加入设计浓度的原花青素培养液，经过相应的培养时间后，再与含有不同浓度过氧化氢的培养液共同作用。流式细胞仪检测葡萄籽原花青素对细胞
贾智艳1，*，
马学华2，
（ 1、哈尔滨医科大学附属一院眼科医院，哈尔滨，150001； 2、哈尔滨医科大学附属一院，哈尔滨，150001； ）
摘要： 目的：评价葡萄籽原花青素对过氧化氢导致的人晶状体上皮细胞损伤的保护作用。 方法：对人晶状体上皮细胞进行培养，分别加入设计浓度的原花青素培养液，经过相应的培养时间后，再与含有不同浓度过氧化氢的培养液共同作用。流式细胞仪检测葡萄籽原花青素对细胞内活性氧的影响。用western blot 方法检测各组细胞中p38，p-p38，JNK, p- JNK蛋白的变化。结果：在体外HLE-B3细胞培养状态下，葡萄籽原花青素能有效地降低细胞内活性氧水平。蛋白测定表明，GSPE能够有效抑制p-p38，p- JNK蛋白表达。结论：葡萄籽原花青素能够减少过氧化氢诱导的细胞内活性氧的产生，从而对细胞具有保护作用。葡萄籽原花青素能够有效抑制p-p38，p-JNK蛋白表达，阻断了MAPK细胞信号传导通路，减少氧化损伤的发生。葡萄籽原花青素对过氧化氢诱导的人晶状体上皮细胞氧化损伤具有保护作用。
关键词： 白内障；原花青素；人晶状体上皮细胞；氧自由基；细胞信号传导
Jia Zhiyan1,*， Ma Xuehua2,
（ 1、Department of Ophthalmology, The First Affiliated Hospital of Harbin Medical University, Harbin, P.R. China.150001； 2、The First Affiliated Hospital of Harbin Medical University, Harbin, P.R. China.150001； ）
Abstract： Purpose：Oxidative damage induced by H2O2 treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. The study described herein investigated the protective effect of Grape seed proanthocyanidin extract (GSPE) on H2O2-induced oxidative stress in human lens epithelial B-3 (HLEB-3) cells and the possible molecular mechanism involved.Methods：After HLE-B3 cells grown to 75%-80% confluence, cells were pretreated with 2 GSPE. After incubation for the indicated time, cells were treated with H2O2 in combination. The ROS generation were detected by flow cytometric analysis. Expression of mitogen activated protein kinase (MAPK) proteins were measured by western blot.Results：GSPE clearly reduced H2O2 induced cell reactive oxygen species (ROS) generation and protected HLEB-3 cells from H2O2 induced oxidative damage. GSPE depressed H2O2-induced phosphorylation of the p38 and c-Jun N-terminal kinase (JNK) proteins of the MAPK family at various time points studied.Conclusions：GSPE could be useful in attenuation of H2O2-induced oxidative stress and the activation of MAPK signaling in HLE-B3 cells, which suggests that GSPE has a potential protective effect against cataractogenesis.
Keywords： cataract; proanthocyanidi; HLE-B3；reactive oxygen species；cellular signaling pathways
作者简介： 贾智艳（1976-），女，副教授，主要研究方向：小儿眼科，斜视弱视
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